Expression profiling and sequence diversity of novel DREB genes from common bean (Phaseolus vulgaris L.) and their association with drought-related traits

Common bean is a major dietary component in several countries, but its productivity is negatively affected by abiotic stresses. Dissecting candidate genes involved in abiotic stress tolerance is a paramount step toward the improvement of common bean performance under such constraints. Thereby, this thesis presents a systematic analysis of the DEHYDRATION RESPONSIVE ELEMENT-BINDING (DREB) gene subfamily, which encompasses genes that regulate several processes during stress responses, but with limited information for common bean. First, a series of in silico analyses with sequences retrieved from the P. vulgaris genome on Phytozome supported the categorization of 54 putative PvDREB genes distributed within six phylogenetic subgroups (A-1 to A-6), along the 11 chromosomes. Second, we cloned four novel PvDREB genes and determined their inducibility-factors, including the dehydration-, salinity- and cold-inducible genes PvDREB1F and PvDREB5A, and the dehydration- and cold-inducible genes PvDREB2A and PvDREB6B. Afterwards, nucleotide polymorphisms were searched through Sanger sequencing along those genes, revealing a high number of single nucleotide polymorphisms within PvDREB6B by the comparison of Mesoamerican and Andean genotypes. The nomenclature of PvDREB6B is discussed in details. Furthermore, we used the BARCBean6K_3 SNP platform to identify and genotype the closest SNP to each one of the 54 PvDREB genes. We selected PvDREB6B for a broader study encompassing a collection of wild common bean accessions of Mesoamerican origin. The population structure of the wild beans was accessed using sequence polymorphisms of PvDREB6B. The genetic clusters were partially associated with variation in latitude, altitude, precipitation and temperature throughout the areas such beans are distributed. With an emphasis on drought stress, an adapted tube-screening method in greenhouse conditions enabled the phenotyping of several drought-related traits in the wild collection. Interestingly, our data revealed a correlation between root depth, plant height and biomass and the environmental data of the location of the accessions. Correlation was also observed between the population structure determined through PvDREB6B and the environmental data. An association study combining data from the SNP array and DREB polymorphisms enabled the detection of SNP associated with drought-related traits through a compressed mixed linear model (CMLM) analysis. This thesis highlighted important features of DREB genes in common bean, revealing candidates for further strategies aimed at improvement of abiotic stress tolerance, with emphasis on drought tolerance

Fleshing Out and Losing Flesh: The Decomposing Corpse as Narrative Object in American Hardboiled Detective Fiction

This dissertation examines the corpse as an object in and of American hardboiled detective fiction written between 1920 and 1950. I deploy several theoretical frames, including narratology, body-as-text theory, object relations theory, and genre theory, in order to demonstrate the significance of objects, symbols, and things primarily in the clever and crafty work of Dashiell Hammett (1894-1961) and Raymond Chandler (1888-1959), but also touching on the writings of their lesser known accomplices. I construct a literary genealogy of American hardboiled detective fiction originating in the writings of Edgar Allan Poe, compare the contributions of classic or Golden Age detective fiction in England, and describe the socio-economic contexts, particularly the predominance of the “pulps,” that gave birth to the realism of the Hardboiled School. Taking seriously Chandler’s obsession with the art of murder, I engage with how authors pre-empt their readers’ knowledge of the tricks of the trade and manipulate their expectations, as well as discuss the characteristics and effect of the inimitable hardboiled style, its sharpshooting language and deadpan humour. Critical scholarship has rarely addressed the body and figure of the corpse, preferring to focus instead on the machinations of the femme fatale, the performance of masculinity, or the prevalence of violence. I cast new light on the world of hardboiled detective fiction by dissecting the corpse as the object that both motivates and de-composes (or rots away from) the narrative that makes it signify. I treat the corpse as an inanimate object, indifferent to representation, that destabilizes the integrity and self-possession, as well as the ratiocination, of the detective who authors the narrative of how the corpse came to be. The corpse is all deceptive and dangerous surface rather than the container of hidden depths of life and meaning that the detective hopes to uncover and reconstruct. I conclude with a chapter that is both critical denouement and creative writing experiment to reveal the self-reflexive (and at times metafictional) dimensions of hardboiled fiction. My dissertation, too, in the manner of hardboiled fiction, hopes to incriminate my readers as much as enlighten them.

Constituição e modelo de mercado : uso liberal e desuso neoliberal

Tese de doutoramento, Direito (Ciências Jurídico-Políticas), Universidade de Lisboa, Faculdade de Direito, 2016

Hemorragia subaracnoideia (HSA) por rotura de aneurisma cerebral em doentes admitidos no Hospital de Santa Maria (HSM) : análise retrospetiva de 2008 a 2012

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

The EU and ASEAN – In Search of a New Regional Paradigm? EU Centre Singapore Working Paper No. 15, August 2013

The European Union (EU) has been hailed as the most successful model of regional integration thus far, while the Association of Southeast Asian Nations (ASEAN), despite its fair share of critics and doomsayers, has been seen as a relatively successful regional organisation in the developing world. However, both seemed to have arrived at a critical juncture in their respective regional projects. Challenged by recent events, internal and external, and faced with increasing uncertainties and complexities, the EU and ASEAN are forced to re-examine the journey they have taken so far and ponder the road ahead. This paper seeks first to provide an overview of the two parallel processes of regionalism in Europe and Southeast Asia by focusing on the developments of the EU and ASEAN, and dissecting both the external forces and internal dynamics that shape the respective regional processes. It then sketches out some of the global trends likely to impact regional developments in Europe and Asia, and questions if the EU and ASEAN would need a new regional approach or paradigm if they are to maintain their salience and relevance as regional actors.

Higher education scholarships as a soft power tool: an analysis of its role in the EU and Singapore. EU Centre Singapore Working Paper No. 23, March 2015

The European Union and Singapore are vastly different entities, each with its own regional and global priorities and policies. Both actors employ a range of tools and instruments to aid in their foreign policy objectives, including in the projection of their soft power. It is worth analysing and comparing the specific instruments of these two actors’ soft power strategies, including but not limited to their stated objectives and perceived effectiveness. This paper will compare the role of higher education and scholarships in diffusing soft power through a comparative case study of the Erasmus Mundus scholarship program and the Singapore Scholarship administered by the Ministry of Foreign Affairs. It will look at the ways in which these programs have shaped the standing of the actors in diffusing their norms and objectives in the regional and international arena. A comparative analysis of these programs will hopefully provide some insight into the proximity between foreign policy-making and higher education internationalisation. This paper will begin with an overview of the aforementioned programs and related schemes, before dissecting and comparing the intent and the policy-making processes behind these, and concludes with a discussion on the present and future role of higher education as a strategic soft power tool.

Banks and the political economy of the sovereign debt crisis in Italy and Spain. CES Papers - Open Forum #18, 2013-2014

This paper sets out to explain why Spain experienced a full-fledged sovereign debt crisis and had to resort to euroarea financial assistance for its banks, whereas Italy did not. It undertakes a structured comparison, dissecting the sovereign debt crisis into a banking crisis and a balance of payments crisis. It argues that the distinctive features of bank business models and of national banking systems in Italy and Spain have considerable analytical leverage in explaining the different scenarios of the crises in each country. This ‘bank-based’ analysis contributes to the flourishing literature that examines changes in banking with a view to account for the differentiated impact of the global banking crisis first and the sovereign debt crisis in the euroarea later.

Lake sediments from Store Koldewey, Northeast Greenland

A 2.9 m long sedimentary record was studied from a small lake, here referred to as Duck Lake, located at 76°25'N, 18°45'W on Store Koldewey, an elongated island off the coast of Northeast Greenland. The sediments were investigated for their geophysical and biogeochemical characteristics, and for their fossil chironomid assemblages. Organic matter began to accumulate in the lake at 9.1 cal. kyr BP, which provides a minimum age for the deglaciation of the basin. Although the early to mid-Holocene is known as a thermal maximum in East Greenland, organic matter accumulation in the lake remained low during the early Holocene, likely due to late plant immigration and lack of nutrient availability. Organic matter accumulation increased during the middle and late Holocene, when temperatures in East Greenland gradually decreased. Enhanced soil formation probably led to higher nutrient availability and increased production in the lake. Chironomids are abundant throughout the record after 9.1 cal. kyr BP and seem to react sensitively to changes in temperature and nutrient availability. It is concluded that relative temperature reconstructions based on biogeochemical data have to be regarded critically, particularly in the period shortly after deglaciation when nutrient availability was low. Chironomids may be a suitable tool for climatic reconstructions even in those high arctic environments. However, a better understanding of the ecology of chironomids under these extreme conditions is needed.

Salivary Gland and Tooth Abnormalities Massively Increase Caries Susceptibility in A Mouse Model of Cleft Lip/Palate

Thesis (Ph.D.)--University of Washington, 2016-04

Haemodynamic assessment following inferior vena cava resection without replacement

Background: Treatment of bulky retroperitoneal malignancy may require en bloc resection of the infrarenal inferior vena cava. A number of reconstructive options are available to the surgeon but objective haemodynamic assessment of the peripheral venous system following resection without replacement is lacking. The aim of the present paper was thus to determine the symptomatic and haemodynamic effects of not reconstructing the resected infrarenal inferior vena cava. Methods: A retrospective descriptive study was carried out at Princess Alexandra Hospital in Queensland. Five patients underwent resection of the thrombosed infrarenal inferior vena cava as part of retroperitoneal lymph node dissection for testicular cancer (n = 3), radical nephrectomy for renal cell carcinoma (n = 1) and thrombosed inferior vena cava aneurysm (n = 1). Clinical effects were determined via the modified venous clinical severity score and venous disability score. Haemodynamic data were obtained postoperatively using venous duplex ultrasound and air plethysmography. Results: None of the present patients scored >2 (out of 30) on the modified venous clinical severity score or >1 (out of 3) on the venous disability score. Haemodynamic studies showed only minor abnormalities. Conclusions: Not reconstructing the resected thrombosed infrarenal inferior vena cava results in minor signs and symptoms of peripheral venous hypertension and only minor abnormalities on haemodynamic assessment.

The molecular genetics of breast cancer: The contribution of comparative genomic hybridization

Comparative genomic hybridization (CGH) has been the technique of choice over the last 10 years for mapping DNA copy number changes in human tumors. Here we review the literature to demonstrate how CGH has contributed to the comprehension of molecular aspects of breast tumorigenesis. At least two distinct molecular pathways of breast cancer have been characterized that show a strong correlation with histological grade. It seems that grade I invasive ductal carcinomas (IDCs) arise from well-differentiated ductal carcinoma in situ (DCIS), whereas grade III IDCs come from poorly differentiated DCIS. In addition, dedifferentiation from a low- to a high-grade breast cancer has proven an unlikely phenomenon. CGH has been instrumental in dissecting distinct molecular pathways toward breast malignancy and in establishing a direct relationship between genotype and clinical pathological features. (C) 2005 Elsevier GrnbH. All rights reserved.

Lessons from tomographic studies of the mammalian Golgi

Basic structure studies of the biosynthetic machinery of the cell by electron microscopy (EM) have underpinned much of our fundamental knowledge in the areas of molecular cell biology and membrane traffic. Driven by our collective desire to understand how changes in the complex and dynamic structure of this enigmatic organelle relate to its pivotal roles in the cell, the comparatively high-resolution glimpses of the Golgi and other compartments of the secretory pathway offered to us through EM have helped to inspire the development and application of some of our most informative, complimentary (molecular, biochemical and genetic) approaches. Even so, no one has yet even come close to relating the basic molecular mechanisms of transport, through and from the Golgi, to its ultrastructure, to everybody's satisfaction. Over the past decade, EM tomography has afforded new insights into structure -function relationships of the Golgi and provoked a re-evaluation of older paradigms. By providing a set of tools for structurally dissecting cells at high-resolution in three-dimensions (3D), EM tomography has emerged as a method for studying molecular cell biology in situ. As we move rapidly toward the establishment of molecular atlases of organelles through advances in proteomics and genomics, tomographic studies of the Golgi offer the tantalizing possibility that one day, we will be able to map the spatio-temporal coordinates of Golgi-related proteins and lipids accurately in the context of 4D cellular space. (c) 2005 Elsevier B.V. All rights reserved.

Lifestyle still predicts mortality in older men with established vascular disease

Background. It is uncertain whether accepted associations between health behaviors and mortality are pertinent to elderly people. No previous studies have examined the patterns of lifestyle in elderly men with and without clinically evident vascular disease by using a lifestyle score to predict survival. Methods. We measured prevalence of a healthy lifestyle (four or more healthy behaviors out of eight) and examined survival in 11,745 men aged 65-83 years participating in a randomized population-based trial of screening for abdominal aortic aneurysm in Perth, Western Australia. After stratifying participants into five groups according to history and symptoms of vascular disease, we compared survival of men in each subgroup with that of 'healthy' men with no history or symptoms of vascular disease. Results. Invitations to screening produced a corrected response of 70.5%. After adjusting for age and place of birth, having an unhealthy lifestyle was associated with an increase of 20% in the likelihood of death from any cause within 5 years (95% CI: 10-30%). This pattern was consistently evident across subgroups defined by history of vascular disease, but was less evident for deaths from vascular disease. Conclusions. Our results highlight the importance of maintaining a healthy lifestyle through to old age, regardless of history of vascular disease. (c) 2005 Elsevier Inc. All rights reserved.

Antifibrotic activity of an inhibitor of group IIA secretory phospholipase A(2) in young spontaneously hypertensive rats

The development of fibrosis in the chronically hypertensive heart is associated with infiltration of inflammatory cells and cardiac hypertrophy. In this study, an inhibitor of the proinflammatory enzyme, group IIA human secretory phospholipase A(2) (sPLA(2)-IIA), has been found to prevent collagen deposition as an important component of cardiovascular remodeling in a rat model of developing chronic hypertension. Daily treatment of young male spontaneously hypertensive rats (SHR) with an sPLA2-IIA inhibitor (KH064, 5-(4-benzyloxyphenyl)-4S-(phenyl-heptanoylamino)-pentanoic acid, 5 mg/kg/day p.o.) prevented increases in the content of perivascular,(SHR 20.6 +/- 0.9%, n = 5; SHR+KH064 14.0 +/- 1.2%, n = 5) and interstitial (SHR 7.9 +/- 0.3%, n = 6; SHR+KH064 5.4 +/- 0.7%, n = 6) collagen in the left ventricle of rat hearts, but did not affect numbers of infiltrating monocytes/macrophages, left ventricular hypertrophy (SHR 2.88 +/- 0.08, n = 12; SHR+KH064 3.09 +/- 0.08 mg/g body weight, n = 9), increased systolic blood pressure, or thoracic aortic responses. This selective antifibrotic activity suggests that sPLA2-IIA may have an important but specific role in cardiac fibrosis, and that its inhibitors could be useful in dissecting molecular pathways leading to fibrotic conditions.

Studying phenotypic evolution using multivariate quantitative genetics

Quantitative genetics provides a powerful framework for studying phenotypic evolution and the evolution of adaptive genetic variation. Central to the approach is G, the matrix of additive genetic variances and covariances. G summarizes the genetic basis of the traits and can be used to predict the phenotypic response to multivariate selection or to drift. Recent analytical and computational advances have improved both the power and the accessibility of the necessary multivariate statistics. It is now possible to study the relationships between G and other evolutionary parameters, such as those describing the mutational input, the shape and orientation of the adaptive landscape, and the phenotypic divergence among populations. At the same time, we are moving towards a greater understanding of how the genetic variation summarized by G evolves. Computer simulations of the evolution of G, innovations in matrix comparison methods, and rapid development of powerful molecular genetic tools have all opened the way for dissecting the interaction between allelic variation and evolutionary process. Here I discuss some current uses of G, problems with the application of these approaches, and identify avenues for future research.